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In-Depth Characterization of Debranching Type I Pullulanase from Priestia koreensis HL12 as Potential Biocatalyst for Starch Saccharification and Modification
In-Depth Characterization of Debranching Type I Pullulanase from Priestia koreensis HL12 as Potential Biocatalyst for Starch Saccharification and Modification
Author
Prongjit D, Lekakarn H, Bunterngsook B, Aiewviriyasakul K, Sritusnee W, Arunrattanamook N, Champreda V
Thammasat University; National Science & Technology Development Agency - Thailand; National Center Genetic Engineering & Biotechnology (BIOTEC)
Type
Article
Source Title
CATALYSTS
Year
2022
Volume
12
Issue
1
Open Access
gold
Publisher
MDPI
DOI
10.3390/catal12091014
Format
PDF
Abstract
Pullulanase is an effective starch debranching enzyme widely used in starch saccharification and modification. In this work, the biochemical characteristics and potential application of a new type I pullulanase from Priestia koreensis HL12 (HL12Pul) were evaluated and reported for the first time. Through in-depth evolutionary analysis, HL12Pul was classified as type I pullulanase belonging to glycoside hydrolase family 13, subfamily 14 (GH13_14). HL12Pul comprises multi-domains architecture, including two carbohydrate-binding domains, CBM68 and CBM48, at the N-terminus, the TIM barrel structure of glycoside hydrolase family 13 (GH13) and C-domain. Based on sequence analysis and experimental cleavage profile, HL12Pul specifically hydrolyzes only alpha-1,6 glycosidic linkage-rich substrates. The enzyme optimally works at 40 degrees C, pH 6.0, with the maximum specific activity of 181.14 +/- 3.55 U/mg protein and catalytic efficiency (k(cat)/K-m) of 49.39 mL/mg center dot s toward pullulan. In addition, HL12Pul worked in synergy with raw starch-degrading alpha-amylase, promoting raw cassava starch hydrolysis and increasing the sugar yield by 2.9-fold in comparison to the alpha-amylase alone in a short reaction time. Furthermore, HL12Pul effectively produces type III-resistant starch (RSIII) from cassava starch with a production yield of 70%. These indicate that HL12Pul has the potential as a biocatalyst for starch saccharification and modification.
Thailand Graduate Institute of Science and Technology (TGIST), National Science and Technology Development Agency [SCA-CO-256312209-TH]; Thammasat University Research Fund [TUFT 042/2563]; Agricultural Research Development Agency [HRD65050090]
License
CC BY
Rights
Authors
Publication Source
WOS
Document
In-Depth Characterization of Debranching Type I Pullulanase from Priestia koreensis HL12 as Potential Biocatalyst for Starch Saccharification and Modification